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GFP和RFP報(bào)道基因在人體不同細(xì)胞中轉(zhuǎn)導(dǎo)率的比較研究

Transduction Efficiency Comparison of GFP and RFP Reporter Gene in Human Cell Lines of hMSC, Eahy926 and A549

作者: 羅毅  辛毅  張穎  汪勁松  王世強(qiáng)  李景超  高飛  黃益民 
單位:首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院-北京市心肺血管疾病研究所(北京100029)
關(guān)鍵詞: 綠色熒光蛋白(GFP);紅色熒光蛋白(  RFP);轉(zhuǎn)導(dǎo)率 
分類(lèi)號(hào):
出版年·卷·期(頁(yè)碼):2011·30·1(20-25)
摘要:

目的 比較慢病毒載體介導(dǎo)的綠色熒光蛋白(GFP)和紅色熒光蛋白(RFP)報(bào)道基因在人體不同細(xì)胞中的轉(zhuǎn)導(dǎo)率,為再生醫(yī)學(xué)和組織工程學(xué)研究中報(bào)道基因的選擇提供依據(jù)。方法  將構(gòu)建的GFP和RFP慢病毒載體分別轉(zhuǎn)染人骨髓間充質(zhì)干細(xì)胞(hMSC)、人臍靜脈內(nèi)皮細(xì)胞株(Eahy926)和人肺泡上皮細(xì)胞株(A549)。4d后,用流式細(xì)胞儀檢測(cè)GFP和RFP的轉(zhuǎn)導(dǎo)率,并利用激光共聚焦顯微鏡觀察GFP和RFP的表達(dá)特征。 結(jié)果 GFP和RFP在hMSC、Eahy926和A549三種細(xì)胞間的轉(zhuǎn)導(dǎo)率均有顯著性差異(P﹤0.01);在hMSC或Eahy926或A549的同種細(xì)胞中GFP和RFP的轉(zhuǎn)導(dǎo)率也有顯著性差異(P﹤0.01);RFP在部分Eahy926和A549細(xì)胞局部高表達(dá)。結(jié)論  慢病毒載體介導(dǎo)的GFP和RFP報(bào)道基因在人體不同細(xì)胞中的轉(zhuǎn)導(dǎo)率明顯不同。

Objective  By comparing the transduction efficiencies of GFP and RFP reporter genes in human cell lines hMSC, Eahy926 and A549, we provide the choice of reporter genes in the regeneration medicine and the organization engineering researches. Methods  Human cell lines of hMSC, Eahy926 and A549 in vitro were respectively infected with lentiviral vectors encoding GFP and RFP reporter gene at MOI 100. After 4 days of transfection, the transduction efficiencies of GFP and RFP in each cell line were evaluated byflow cytometry. The expressive features of GFP and RFP reporters in cells were observed with confocal microscopy.  Results  We found that the transduction efficiencies of either GFP or RFP among three cell lines were very significantly difference (P<0.01). The transduction efficiencies of GFP and RFP among same cell line also was markedly varied and the difference was statistical significant (P<0.01). The RFP was concentrated expression at the subcellular localization in small population cells of Eahy926 and A549 strain. Conclusion The transduction efficiencies of GFP and RFP were evidently different among hMSC, Eahy926 and A549 cell lines.
 

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